Protein Tagging of GPCR
Creative Biolabs provides high-quality protein tagging services for both GPCR research and pharmaceutical use. Our experienced scientists can help our customers choose protein tags that are right for their experiments and downstream applications.
Introduction of Protein Tagging
Protein tagging is an important aspect of protein analysis, which commonly refers to the fusion expression of a polypeptide or protein with target protein using recombinant DNA technology. A variety of protein tags with different functions have been successively developed and applied in various applications. Most protein tags can be used for protein expression, purification, detection and tracking using such techniques as immunoassays, pull-down assays, Western blotting and ELISA. Certain tags may be used for subcellular localization, protein interaction and functional study.
G protein-coupled receptors (GPCRs), the largest class of membrane proteins in the human genome, are highly relevant to various diseases. GPCR research has always been the forefront and center of drug development and precision medicine. Protein tagging can facilitate a wide variety of studies on their structure and function. However, the nature of the protein tag and its location within the receptor need to be considered carefully.
Figure 1. Some potential applications of epitope-tagging. (Huang Y & Willars G B., 2011)
Overview of Protein Tags
Explore the options below. We can choose the best tag for your specific application.
| Category | Description |
|---|---|
| Affinity tags | Affinity tags added to either the N-terminal or C-terminal of the target protein can facilitate purification from its original biological source using affinity techniques. |
| Solubilization tags | Solubilization tags can be used to assist in proper folding of proteins and keep them from precipitating. |
| Chromatography tags | Chromatography tags can be used to alter chromatographic properties of target proteins. Different resolution can be afforded by particular separation technique. |
| Epitope tags | Epitope tags refers to small antigenic peptides usually derived from viral genes with high-affinity to antibodies. These tags are applicable for protein purification, protein expression and subcellular localization. |
| Fluorescence tags | Fluorescence tags can be used to realize visualization of target proteins. GFP and its variants are commonly used. |
Commonly Used Protein Tags
| Tags | Sequence/Origin | Tags | Sequence/Origin |
|---|---|---|---|
| HA- | YPYDVPDYA | Twin-Strep-tag | SAWSHPQFEKGGGSGGG SGGSAWSHPQFEK |
| Poly-His- | nH (n = 2-12, usually 6) | CBP- | KRRWKKNFIAVSAANRFKKI SSSGAL |
| GST- | Glutathione-S-transferase | Chitin-binding tag | TNPGVSAWQVNTAYTAGQL VTYNGKTYKCLQPHTSLAG WEPSNVPALWQLQ |
| Poly-Arg- | nR (n = 5-6, usually 5) | HAT- | KDHLIHNVHKEFHAHAHNK |
| FLAG- | DYKDDDDK | BAP- | GLNDIFEAQKIEWHE |
| SBR- | MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP | Terbium-binding protein | DDDWDDDWDDDWDDD |
| Strep-tag | WRHPQFGG | GFP | Aequorea victoria (26.9 kDa) |
| S- | KETAAAKFERQHMDS | Halo tag | Rhodococcus (33 kDa) |
| MYC | EQKLISEEDL | NusA | 495 amino acids |
| MBP | AEEGKLVIW | TrxA | Escherichia coli. (12 kDa) |
| c-Myc | EQKLISEEDL | Fh8 | Fasciola hepatica (8 kDa) |
| Avi- | GLNDIFEAQKIEWHE | SUMO | 100 amino acids |
| Strep-tag II | WSHPQFEK | SNAP | O6-alkylguanine-DNA-alkylransferase |
Creative Biolabs is always ready to provide the best services to each of our customers. We can help you choose the appropriate labeling strategies and offer the optimized protocols.
If you are interested in our services or have any specific requirements, please feel free to contact us for more details.
References
- Huang Y & Willars G B. Generation of epitope-tagged GPCRs. Receptor Signal Transduction Protocols. Humana Press, Totowa, NJ, 2011: 53-84.
- Lotze J, et al. Peptide-tags for site-specific protein labelling in vitro and in vivo. Molecular BioSystems, 2016, 12(6): 1731-1745.